Library
The following reference materials are available for researchers to borrow:
- Methods in Cell Biology, Volume 75: Cytometry 4th Edition. New Developments. (2004) Darzynkiewick, Roederer and Tanke (eds)
- Current Protocols in Flow Cytometry
- Course Notebook - 27th Annual Course in Flow Cytometry
PROTOCOLS
These flow protocols have been validated by members of the Flow Core.
Software Guides
Preparing a FACSDiva Experiment
Sort Forms and Instrument Issues
Flow Core Database - fill out a sort form or report an issue here
Sort Form Information - information about filling in the sort form
Troubleshooting and After Hours
Peak 6 Bead Information
The reference bead that is used in the facility is from Spherotech. We use the Rainbow Calibration Particles, 6th peak of 8. The part number for this product is RCP-30-5A-6 and the current lot that we use is AC01. With the recent change of bead lots, we also are supplying this excel file to illustrate the changes between the previous bead lot (AB02) and the new lot (AC01).
Connecting to the Transfer space
Double click the "shortcut to transfer" icon on the desktop. An explorer window with a folder for each PI will open. NOTE - you will not be able to read the contents of any of these folders, even your own. These folders are write only from the instruments. As before, drag the folders you want to transfer from the FCS folder to your group folder. If you wish to make sure a transfer happened, try it again. You will be told if the file is already there. The folder will report as empty, since it is not readable.
To recover your data on your lab computer:
For PC:
To connect to RDS01 in Windows 7:
Click the "Start" button then in the "Search programs and files" field
that appears type "\\rds01" and hit the "Enter" key on your keyboard.
If you had Windows remember your credentials then you will need to delete
those. You can do this by clicking the "Start" button and then in the
"Search programs and files" field that appears type "credential manager".
You should see a suggested icon of a safe appear in the list above the
search field that says "Credential Manager". Click this, it will open a
new window showing your saved credentials. Under "Windows Credentials"
delete anything related to RDS01. If you are in doubt feel free to delete
a Windows Credential, the worst case scenario is you will have to re-input
your password for something else.
To connect to RDS01 in Windows XP:
Click the "Start" button and choose "Run". Clear any text that may be in
the Run box and type "\\rds01". Click "OK".
For Mac:
Click the "Go" menu in Finder, choose "Connect to Server". Clear out
anything that is in the "Server Address" box and type "smb://rds01". Click
"Connect". If you are prompted for a username and password use your URMC
network credentials (what you would use for webmail or to log onto a
Windows computer here) instead of what you used before. You should be
presented with a list of shares, please choose the one corresponding to
your PI's name.
REMEMBER!!
Data will be automatically removed from the transfer partition after two weeks.
Connecting to the Library Share
This is done similarly to the Transfer space. Here we have shared documents that are available to all registered users. This includes past lectures, IDEAS software, and other goodies.
For PC:
Start Menu > Run...
\\rds01\library
For Mac:
Finder >Go > Connect to server
smb://rds01/library
Additional FlowJo Server information
PDF available (behind the firewall)
Current Instrument Baseline
Grant Information
The URMC Flow Cytometry Core facility (FCC), under the direction of Timothy Bushnell, Ph.D., is a full service facility with approximately 1100 sq. feet of newly renovated lab space, employing a staff of 6. The facility currently has a range of digital flow cytometers including a 2-laser, 4-color Accuri C6, a 3-laser, 8-color BD-FACSCanto-II, two 3-laser, 12-color BD LSR-II’s, and two 4-laser, 18-color BD LSR-II’s. Additionally, the facility is equipped with a 3-laser, 13-color FACSAria-Iiu, a 4-laser, 18-color FACSAria-II, both of which are equipped with aerosol management systems, automatic cell deposition units, and are housed within Baker Bio-Protect-III biosafety hoods. The facility also has a 3-laser, 6-channel IS100 and 4-laser, 12-channel ImagestreamX imaging flow cytometers from Amnis. The facility has three dedicated analysis workstations for investigators with standard flow cytometric analysis software (Flowjo, IDEAS, Modfit). All data generated in the FCC is archived onto a 10-terabyte server with daily tape backup.
The FCC has established a series robust QA/QC protocols and procedures that are designed to ensure the instruments are performing optimally. These include standard quality control measurements, as recommended by the instrument manufacturers. Additional detailed instrumentation characterization and QC are performed based on best practices in the field. All QA/QC information is available to investigators and instrumentation characterization data is available on the FCC’s website.
All investigators go through a robust training program to allow them run the analyzers independently, allowing for experienced users to have 24/7 access. All booking is done via an on-line calendar system. All sorting is done by the FCC staff, and is available to investigators from ~10 am till 9:30 pm daily.
Recent Publications
- Enos, M., Bancos, S., Bushnell, T., and Crispe, I.N. (2008) E2F4 modulates differentiation and gene expression in hematopoietic progenitor cells during commitment to the lymphoid lineage J Immunol 180:3699-707
- McGrath, K.E., Kingsley, P.D., Koniski, A..D., Porter, R.L., Bushnell, T.P., and Palis, J.(2007) Enucleation of primitive erythroid cells generates a transient population of "pyrenocytes" in the mammalian fetus. Blood 111:2409-17
- Guzman, M.L., Li, X., Corbett, C.A., Rossi, R.M., Bushnell, T., Liesveld J.L., Herbet, J., Young, F., and Jordan C.T. (2007) Rapid and selective death of leukemia stem and progenitor cells induced by the compound 4-benzyl, 2-methyl, 1,2,4-thiadiazolidine, 3,5 dione (TDZD-8). Blood 110:4436-44
- Ryan, E.P., Bushnell, T.P., Friedman, A.E., Rahman, I., and Phipps, R.P. (2007) Cyclooxygenase-2 selective inhibition reduces B lymphoma survival by inducing oxidative stress and decreasing intracellular glutathione Cancer Immunol Immunother 57:347-58
- Neering, S.J., Bushnell, T., Sozer, S., Ashton, J., Rossi, R.M., Wang, P.-Y., Bell, D.R., Heinrich, D., Bottaro, A, and Jordan C.T. (2007) Leukemia stem cells in a genetically defined murine model of blast crisis CML. Blood 110:2578-85.
- Graf, B., Bushnell, T., and Miller , J. (2007) LFA-1-mediated T cell costimulation through increased localization of TCR/class II complexes to the cSMAC and exclusion of CD45 from the immunological synapse. J Immunol., 179:1616-24
- Neff-LaFord, H., Teske, S., Bushnell, T.P., and Lawrence, B.P. (2007) Aryl hydrocarbon receptor activation during influenza virus infection unveils a novel pathway of IFN-a production by phagocytic cells J Immunol 179:247-55
- Kingsley, PD, Malik J, Emerson RL, Bushnell TP, McGrath KE, Bloedorn LA, Bulger M, and Palis J (2006) “Maturational” globin switching in primary primitive erythroid cells. Blood 107:1665-1672
Upcoming Events
PPMS going live April 1st. Please make sure you are familiar with how it works.
Training Lecture (Accuri, LSR, Canto) - MC 3-6232
May 27, June 10 and 17th @ 4PM
Lecture series
next lecture coming soon!
Spectral Viewers
Spectra Viewer (via Invitrogen)
Spectra Viewer (via BD Biosciences)
Spectra Viewer (via EBioscience)
Instrument Guidelines
Comments about the Flow Core?
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